REST Inactivation and Coexpression of ASCL1 and POU3F4 Are Necessary for the Complete Transformation of RB1/TP53-Inactivated Lung Adenocarcinoma into Neuroendocrine Carcinoma
REST Inactivation and Coexpression of ASCL1 and POU3F4 Are Necessary for the Complete Transformation of RB1/TP53-Inactivated Lung Adenocarcinoma into Neuroendocrine Carcinoma
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DOI：
10.1016/j.ajpath.2022.03.007
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发表时间：
2022-06-06
影响因子：
6
通讯作者：
Yazawa, Takuya
中科院分区：
文献类型：
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作者：
Masawa, Meitetsu;Sato-Yazawa, Hanako;Yazawa, Takuya
尽管最近的报道揭示了RB1和TP53的失活在肺腺癌向神经内分泌癌（NEC）转化中的重要性，但完全转化为NEC的要求尚未阐明。为了研究与RB1/TP53失活相关的特征的改变并确定转化成NEC细胞的要求，建立了RB1/TP53双敲除的A549肺腺癌细胞，并进行了REST的额外敲除和ASCL1和POU 3类同源框转录因子（TF）的转染。在RB1/TP53双基因敲除的A549细胞中，超过60个在神经细胞中大量表达的基因和几个与上皮向间质转化相关的基因被上调。虽然嗜铬粒蛋白A和突触素的表达诱导额外的敲除REST（模拟大多数NEC的状态），另一种神经内分泌标志物，CD56和前神经TF的表达没有诱导。然而，ASCL1和POU3F4在RB1/TP53/REST三基因敲除的A549细胞中的共表达不仅诱导了CD56的表达，而且还诱导了其他前神经TF（NEUROD1和胰岛素瘤相关1）的表达，并诱导了NEC样形态。这些发现表明，RB1和TP53的失活诱导肺腺癌转化为NEC所必需的状态，REST的进一步失活以及ASCL1和POU3F4的共表达是完全转化为NEC的触发因素。
Although recent reports have revealed the importance of the inactivation of both RB1 and TP53 in the transformation from lung adenocarcinoma into neuroendocrine carcinoma (NEC), the requirements for complete transformation into NEC have not been elucidated. To investigate alterations in the characteristics associated with the inactivation of RB1/TP53 and define the requirements for transformation into NEC cells, RB1/TP53 double-knockout A549 lung adenocarcinoma cells were established, and additional knockout of REST and transfection of ASCL1 and POU class 3 homeobox transcription factors (TFs) was conducted. More than 60 genes that are abundantly expressed in neural cells and several genes associated with epithelial-to-mesenchymal transition were up-regulated in RB1/TP53 double knockout A549 cells. Although the expression of chromogranin A and synaptophysin was induced by additional knockout of REST (which mimics the status of most NECs), the expression of another neuroendocrine marker, CD56, and proneural TFs was not induced. However, coexpression of ASCL1 and POU3F4 in RB1/TP53/REST triple-knockout A549 cells induced the expression of not only CD56 but also other proneural TFs (NEUROD1 and insulinoma-associated 1) and induced NEC-like morphology. These findings suggest that the inactivation of RB1 and TP53 induces a state necessary for the transformation of lung adenocarcinoma into NEC and that further inactivation of REST and coexpression of ASCL1 and POU3F4 are the triggers for complete transformation into NEC.